DEAD-box ATPases are global regulators of phase-separated organelles.

The ability of proteins and nucleic acids to undergo liquid-liquid phase separation has recently emerged as an important molecular principle of how cells rapidly and reversibly compartmentalize their components into membrane-less organelles such as the nucleolus, processing bodies or stress granules1,2. How the assembly and turnover of these organelles are controlled, and how these biological condensates selectively recruit or release components are poorly understood. Here we show that members of the large and highly abundant family of RNA-dependent DEAD-box ATPases (DDXs)3 are regulators of RNA-containing phase-separated organelles in prokaryotes and eukaryotes. Using in vitro reconstitution and in vivo experiments, we demonstrate that DDXs promote phase separation in their ATP-bound form, whereas ATP hydrolysis induces compartment turnover and release of RNA. This mechanism of membrane-less organelle regulation reveals a principle of cellular organization that is conserved from bacteria to humans. Furthermore, we show that DDXs control RNA flux into and out of phase-separated organelles, and thus propose that a cellular network of dynamic, DDX-controlled compartments establishes biochemical reaction centres that provide cells with spatial and temporal control of various RNA-processing steps, which could regulate the composition and fate of ribonucleoprotein particles.

Sex differences in cardiologic medication provision for adults with coronary heart disease: an analysis of health claims data from 2018 to 2020 in Saxony-Anhalt, Germany.

BACKGROUND: Coronary heart diseases (CHDs) have experienced the largest increase worldwide as a cause of death, accounting for 16% of all deaths. In Saxony-Anhalt, a federal state in Germany, both CHD morbidity and acute myocardial infarction mortality rates are particularly high. Several risk factors associated with CHDs have been studied in Saxony-Anhalt, but sex differences in service use and medication have not been investigated. This study therefore aimed to investigate sex differences in the quality and quantity of cardiological care provided to adults with CHD. METHODS: This study used health claims data from 2018 to 2020 to analyse the utilisation of healthcare services and adherence to medication-related guideline recommendations in primary and specialist care. The sample included 133,661 individuals with CHD from a major statutory health insurance company (Germany). RESULTS: Almost all CHD patients (> 99%) received continuous primary care. Continuous cardiologist utilisation was lower for females than for males, with 15.0% and 22.2%, respectively, and sporadic utilisation showed greater differences, with 33.5% of females and 43.4% of males seeking sporadic cardiologist consultations. Additionally, 43.1% of the identified CHD patients participated in disease management programmes (DMPs). The study also examined the impact of DMP participation and cardiologist care on medication uptake and revealed that sex differences in medication uptake, except for statin use, were mitigated by these factors. Statins were prescribed to 42.9% of the CHD patients eligible for statin prescription in accordance with the QiSA indicator for statin prescription eligibility. However, there were significant sex differences in statin utilisation. Female CHD patients were less likely to use statins (35.2%) than male CHD patients were (50.1%). The difference in statin utilisation persisted after adjustment for DMP participation and cardiologist consultation. CONCLUSIONS: This study highlights sex differences in the utilisation of cardiological healthcare services for patients with CHD in the Saxony-Anhalt cohort. These findings underscore the continuing need for interventions to reduce sex inequalities in accessing healthcare and providing health care for patients with CHD. Factors at the health care system, patient, and physician levels should be further investigated to eventually improve statin prescription in people with CHD, especially women.

[Development Of Long-Term Care Dependency And Utilisation Of Long-Term Care Services From 2017 To 2022 In Germany, Saxony-Anhalt: Analysis Of Health Insurance Data]. / Entwicklung der Pflegebedürftigkeit und Inanspruchnahme pflegerischer Leistungen von 2017 bis 2022 auf Basis der Routinedaten der AOK Sachsen-Anhalt.

BACKGROUND: Saxony-Anhalt has the highest old-age dependency ratio among the German federal states. The proportion of long-term care-dependent people can be expected to increase in the future, given the aging of the population. The SARS-CoV-2 pandemic has influenced nursing care, reduced the utilisation of services and led to changes in care arrangements. The aim of this study was to analyse the development of long-term care-dependency in Saxony-Anhalt, Germany, taking into account the legal changes, the pandemic and the development after the lifting of the contact ban. METHOD: The analysis uses aggregated and anonymised health claims data from people insured with AOK Saxony-Anhalt from 2017 to 2022 in need of long-term care. Changes over time in the use of various care services were analysed. RESULTS: In 2017, 64,591 people insured in the statutory health insurance AOK Saxony-Anhalt (AOK ST) were in need of long-term care. The prevalence increased continuously throughout the observation years. For the lowest level of care (minor impairment of independence), the most significant and strongest increase was recorded in the first years after its introduction from 2017-2019. The majority of people in need of long-term care (41-44%) were in the category of care level 2 (considerable impairment of independence). Overall, the proportion of cash benefits has risen steadily since 2017, from 40.5% to 50.3% in 2022. At the same time, the proportion of long-term care in nursing homes decreased at all care levels. The trend toward less nursing home care has become stronger since 2017 and particularly since the onset of the SARS-CoV-2 pandemic in 2020. CONCLUSION: Since the introduction of the new definition of the need for long-term care in Germany, the analysis shows an increase in the number of people in need of long-term care, with those with considerable impairment of independence making up the largest proportion. There has been a significant increase in the number of people receiving cash benefits and a decline in nursing home care. Compared to the nationwide data of health claims, data show a greater use of nursing services in home-based arrangements. The analysis during the COVID-19 pandemic shows a trend towards home care, but no fundamental change. Special situations such as contact restrictions may have influenced the use of care services. Future research should examine the design of home care arrangements and the needs of those affected in greater detail.

[Utilization of pre-hospital emergency medical services: Secondary data analysis of health insurance data from Saxony-Anhalt]. / Inanspruchnahme der bodengebundenen Notfallrettung ­ Sekundärdatenanalyse der Versichertenkohorte AOK Sachsen-Anhalt.

BACKGROUND: Utilization of emergency care is subject to demographic changes. In Germany, there was an increase of 4% to 6% annually between the years 2000 and 2017. Studies revealed that age and regional structures were related to utilization. The aim of this cross-sectional study was to analyze utilization of pre-hospital emergency medical services in Saxony-Anhalt and to identify associated factors. METHOD: Based on data from the AOK Saxony-Anhalt for 2019, transportation with ground ambulances of insured persons living in Saxony-Anhalt were identified and recorded. Insured time of all AOK insured persons living in Saxony-Anhalt was used to determine incidence rates. For the multivariate analysis of associated factors, a Poisson regression model was adapted. RESULTS: 112,575 transportations of patients were identified; an average of 1.53 events per person or 149.6 per 1,000 person years. Inpatient hospital treatment was provided for 53.2% of emergency patients and outpatient hospital treatment for 37.1%. Frequent users represented 14.6% of the events. The incidence rate was highest for older persons and those with care dependency. In addition, the risk of using emergency services was higher among men and in suburban and urban regions (compared with rural regions), even after adjustment. CONCLUSIONS: Utilization is higher in the cohort studied compared to that found in other studies. The high proportion where inpatient hospital treatment is not necessary, as well as the clear association with care dependency and age, offer possible angles of approach for interventions to relieve the burden on emergency services.

Evaluation of two different semi-automated homogenization techniques in microbiological diagnosis of periprosthetic joint infection: disperser vs. bead milling method.

BACKGROUND: In microbiological diagnosis of periprosthetic joint infection (PJI) there is no consensus regarding the most suitable and optimal number of specimens to be cultured or the most effective technique of tissue processing. This comparative study analysed the accuracy of two semi-automated homogenization methods with special focus on the volume and exact origin of each sample. METHODS: We investigated a total of 722 periprosthetic tissue samples. PJI was defined according to the new scoring system for preoperative and intraoperative criteria. We compared the performance of our routinely used single tissue processing by disposable high-frequency disperser with the bead milling method. RESULTS: Eighty patients were included. Among forty classified PJIs, 34 patients yielded positive culture results. In 23 cases (68%) exact concordant results were generated with both techniques. However, in seven cases (20%) processing by the disperser and in four cases (12%) by bead milling provided additional positive samples, but without significant difference since the major definition criteria were met in all cases. The percentage of positive results was influenced by the volume and origin of the tissue samples. Results for small tissue samples tended to be better using the bead milling method. This might lead to improved preoperative arthroscopic diagnosis, as the volume of biopsies is generally limited. Six patients had negative results due to previous antimicrobial therapy. Forty other patients were classified as aseptic failures. Neither procedure resulted in any contamination. CONCLUSION: Both methods enable reliable processing of tissue samples for diagnosis of PJI and are suitable for routine use.

[Dementia Care Nurse - Feasibility study of outreach assistance for people with dementia and their family carers]. / Dementia Care Nurse.

Dementia Care Nurse - Feasibility study of outreach assistance for people with dementia and their family carers Abstract. Background: Case management for dementia is stipulated in the German national dementia strategy. The effectiveness of case management has been investigated many times, yet the results are heterogeneous. On the contrary, the implementation processes of case management concepts have to date rarely been described or evaluated in detail. Aim: The aim of the project was to analyze the implementation of an outreaching assistance for people with dementia and to explore the changes in care, acceptance, and also facilitators and barriers to the implementation. Methods: Between 08 / 2018 and 07 / 2019, outreaching assistance for people with dementia and their family carers was implemented. Quantitative and qualitative data were prospectively collected using semi-standardised interviews in the context of outreach assistance. Results: A total of 113 people with dementia were included in the study, and for the most part family carers could be involved. On average, eight contacts took place over a period of 74 days. The areas of need and support were diverse. The use of support services increased by 19 % after the end of the intervention. Conclusions: The implementation of continuous and processual support for people with dementia and their carers is possible, whereby the structure and procedure should be transparent. A comprehensive orientation and networking is beneficial. The effectiveness of the intervention remains to be investigated in a controlled study.

Sonicate fluid inoculated into blood culture bottles does not improve diagnosis of periprosthetic joint infection caused by anaerobes. A retrospective analysis.

BACKGROUND: In microbiological diagnosis of periprosthetic joint infection (PJI) there is much controversial discussion about culture media and incubation time, especially if anaerobic bacteria are the causative agents. This retrospective analysis was conducted to compare the results obtained by inoculation of sonicate fluid from prosthetic components into BD Bactec blood culture bottles with those obtained by our culture method using sensitive supplemented growth media. METHODS: Twenty-eight cases were included in this study. For definition of PJI, the criteria of the Musculoskeletal Infection Society (MSIS) were considered. The quantity and time to positivity of anaerobes detected in sonicate fluid were monitored both from inoculated supplemented liver thioglycollate broth and anaerobic blood culture bottles. Furthermore, phenotypic testing was performed on the antimicrobial activity within the sonicate fluid. RESULTS: The most frequently isolated microbes were Cutibacterium species, followed by Finegoldia magna, Parvimonas micra, Robinsoniella peoriensis, Clostridium species, Peptoniphilus harei and Slackia exigua. In 24 cases, the microorganisms became detectable within five days (median time 3.2 days) when sonicate fluid was incubated in supplemented liver thioglycollate broth, regardless of whether the patients had taken antimicrobial agents prior to surgery. However, when sonicate fluid was inoculated into anaerobic Bactec bottles, the median time to positivity was 7.4 days and only 12 cases (43%) were correctly identified. Sixteen cases remained negative after 14 days of incubation. CONCLUSION: Depending on the pathogen, incubation of sonicate fluid using blood culture bottles can support diagnosis of PJI but compared with our culture medium it is less efficient if anaerobes are the suspected cause of infection. Microbiological expertise is therefore indispensable to ensure reliable detection of these microorganisms in PJI until a gold standard for laboratory handling of anaerobes has been established.

[Challenges in dementia care at home - The situation at home of a married couple]. / Herausforderungen bei Demenz in den eigenen vier Wänden.

Challenges in dementia care at home - The situation at home of a married couple Abstract. Background: People with dementia and their relatives are faced with major challenges due to complex dementia symptoms. Families need information and counselling in order to find adequate dementia care services tailored to their needs. AIM: This case report's objective is to exemplify the domestic situation of a married couple who is faced with significant challenges within the family and the care system due to the husband's dementia and Parkinson's disease. METHODS: The Dementia Care Nurse project included case monitoring; by means of different assessments relevant information was recorded and the family's situation described. RESULTS: The family's problems and their need for support were multifaceted and entailed reimbursement of costs, application for care services as well as management of challenging behaviours and reduction of the caregiver's psychosocial burden. CONCLUSIONS: The family, particularly the spouse caregiver, was effectively supported in meeting the challenges of dementia, e. g. by drawing on professional services and sorting out entitlement to benefits. From the perspective of the experience in the project, independent counselling structures such as a case management approach are indispenable in order to stabilise the domestic situation.

Die Lebenssituation von Menschen mit Demenz und pflegenden Angehörigen während der Coronavirus-Pandemie Eine qualitative Studie.

The life situation of people with dementia and family carers during the coronavirus pandemic - A qualitative study Abstract. Background: The measures initiated as a result of the coronavirus pandemic have far-reaching consequences for the everyday life of people with dementia and their family carers. Both are usually among those who are the most vulnerable and thus are subject to rigorous restrictions. Their everyday life is made more difficult because care and respite services are currently suspended. In addition, people with dementia have difficulty understanding and implementing the restrictions and hygiene rules. AIMS: This study aims to describe the current life situation of family carers and people with dementia. METHODS: For this purpose, 21 telephone interviews with both family carers and people with dementia have been conducted twice during the spring of 2020 and were subsequently evaluated by a content analysis. RESULTS: The interviewees experienced the situation differently. Especially the social isolation, the higher amount of care, the uncertainty of the situation and the increase in psychological symptoms are described as being stressful. With regard to coping with the situation, discussions are taking place about the support from the social environment, alternative ways of communication, experiences with comparable crises, the stability of formal care and the handling of information. CONCLUSIONS: Family carers and people with dementia feel stressed due to the coronavirus pandemic, but many of them have coping strategies for this special situation. Informal support is a particularly important support mechanism.

Diatrack particle tracking software: Review of applications and performance evaluation.

Object tracking is an instrumental tool supporting studies of cellular trafficking. There are three challenges in object tracking: the identification of targets; the precise determination of their position and boundaries; and the assembly of correct trajectories. This last challenge is particularly relevant when dealing with densely populated images with low signal-to-noise ratios-conditions that are often encountered in applications such as organelle tracking, virus particle tracking or single-molecule imaging. We have developed a set of methods that can handle a wide variety of signal complexities. They are compiled into a free software package called Diatrack. Here we review its main features and utility in a range of applications, providing a survey of the dynamic imaging field together with recommendations for effective use. The performance of our framework is shown to compare favorably to a wide selection of custom-developed algorithms, whether in terms of localization precision, processing speed or correctness of tracks.

Stem-loop RNA labeling can affect nuclear and cytoplasmic mRNA processing.

The binding of sequence-specific RNA-interacting proteins, such as the bacteriophage MS2 or PP7 coat proteins, to their corresponding target sequences has been extremely useful and widely used to visualize single mRNAs in vivo. However, introduction of MS2 stem-loops into yeast mRNAs has recently been shown to lead to the accumulation of RNA fragments, suggesting that the loops impair mRNA decay. This result was questioned, because fragment occurrence was mainly assessed using ensemble methods, and their cellular localization and its implications had not been addressed on a single transcript level. Here, we demonstrate that the introduction of either MS2 stem-loops (MS2SL) or PP7 stem-loops (PP7SL) can affect the processing and subcellular localization of mRNA. We use single-molecule fluorescence in situ hybridization (smFISH) to determine the localization of three independent mRNAs tagged with the stem-loop labeling systems in glucose-rich and glucose starvation conditions. Transcripts containing MS2SL or PP7SL display aberrant localization in both the nucleus and the cytoplasm. These defects are most prominent in glucose starvation conditions, with nuclear mRNA processing being altered and stem-loop fragments abnormally enriching in processing bodies (PBs). The mislocalization of SL-containing RNAs is independent of the presence of the MS2 or PP7 coat protein (MCP or PCP).

Temporal and spatial regulation of mRNA export: Single particle RNA-imaging provides new tools and insights.

The transport of messenger RNAs (mRNAs) from the nucleus to cytoplasm is an essential step in the gene expression program of all eukaryotes. Recent technological advances in the areas of RNA-labeling, microscopy, and sequencing are leading to novel insights about mRNA biogenesis and export. This includes quantitative single molecule imaging (SMI) of RNA molecules in live cells, which is providing knowledge of the spatial and temporal dynamics of the export process. As this information becomes available, it leads to new questions, the reinterpretation of previous findings, and revised models of mRNA export. In this review, we will briefly highlight some of these recent findings and discuss how live cell SMI approaches may be used to further our current understanding of mRNA export and gene expression.

MEMO: multi-experiment mixture model analysis of censored data.

MOTIVATION: The statistical analysis of single-cell data is a challenge in cell biological studies. Tailored statistical models and computational methods are required to resolve the subpopulation structure, i.e. to correctly identify and characterize subpopulations. These approaches also support the unraveling of sources of cell-to-cell variability. Finite mixture models have shown promise, but the available approaches are ill suited to the simultaneous consideration of data from multiple experimental conditions and to censored data. The prevalence and relevance of single-cell data and the lack of suitable computational analytics make automated methods, that are able to deal with the requirements posed by these data, necessary. RESULTS: We present MEMO, a flexible mixture modeling framework that enables the simultaneous, automated analysis of censored and uncensored data acquired under multiple experimental conditions. MEMO is based on maximum-likelihood inference and allows for testing competing hypotheses. MEMO can be applied to a variety of different single-cell data types. We demonstrate the advantages of MEMO by analyzing right and interval censored single-cell microscopy data. Our results show that an examination of censoring and the simultaneous consideration of different experimental conditions are necessary to reveal biologically meaningful subpopulation structures. MEMO allows for a stringent analysis of single-cell data and enables researchers to avoid misinterpretation of censored data. Therefore, MEMO is a valuable asset for all fields that infer the characteristics of populations by looking at single individuals such as cell biology and medicine. AVAILABILITY AND IMPLEMENTATION: MEMO is implemented in MATLAB and freely available via github (https://github.com/MEMO-toolbox/MEMO). CONTACTS: eva-maria.geissen@ist.uni-stuttgart.de or nicole.radde@ist.uni-stuttgart.de SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Individualized support for informal caregivers of people with dementia - effectiveness of the German adaptation of REACH II.

BACKGROUND: Individualized, outreach and structured multicomponent interventions are a promising intervention approach to relieve the burden of informal caregivers of people with dementia. In this study, we adapted and evaluated a multicomponent intervention (Resources for Enhancing Alzheimer's Caregiver Health II, REACH II), which was developed in the USA, to the German health-care system. Therefore the project is called the German adaptation of REACH II (in German: Deutsche Adaptation der REACH II, DE-REACH). METHODS: The effectiveness of DE-REACH was examined in a randomized, controlled trial on 92 informal caregivers of people with dementia. The intervention comprised 12 individual two-weekly sessions (9 at home with the informal caregiver and 3 via telephone) and combined five modules. The reduction of the burden of the informal caregivers was chosen as the primary outcome. RESULTS: The results showed a great stabilizing effect of the intervention on caregiver burden (effect size d = 0.91), that is, comparing pre- and post-measurements the burden decreased very slightly in the intervention group whereas it increased very strongly in the control group. After a three-month follow-up period this effect decreased from a great to a moderate effect. There were also improvements as a result of the intervention in somatization, health-related psychological quality of life and the reaction of the informal caregivers in response to challenging behaviors of the relative with dementia. Moreover, the frequency of challenging behaviors of the affected person itself was reduced in favor of the intervention. CONCLUSION: The findings of this study provide further evidence for the impact of multicomponent support interventions for informal caregivers of people with dementia. CLINICAL TRIAL REGISTRATION: NCT01690117 . Registered September 17, 2012.

Mad1 contribution to spindle assembly checkpoint signalling goes beyond presenting Mad2 at kinetochores.

The spindle assembly checkpoint inhibits anaphase until all chromosomes have become attached to the mitotic spindle. A complex between the checkpoint proteins Mad1 and Mad2 provides a platform for Mad2:Mad2 dimerization at unattached kinetochores, which enables Mad2 to delay anaphase. Here, we show that mutations in Bub1 and within the Mad1 C-terminal domain impair the kinetochore localization of Mad1:Mad2 and abrogate checkpoint activity. Artificial kinetochore recruitment of Mad1 in these mutants co-recruits Mad2; however, the checkpoint remains non-functional. We identify specific mutations within the C-terminal head of Mad1 that impair checkpoint activity without affecting the kinetochore localization of Bub1, Mad1 or Mad2. Hence, Mad1 potentially in conjunction with Bub1 has a crucial role in checkpoint signalling in addition to presenting Mad2.

Mph1 kinetochore localization is crucial and upstream in the hierarchy of spindle assembly checkpoint protein recruitment to kinetochores.

The spindle assembly checkpoint (SAC) blocks entry into anaphase until all chromosomes have stably attached to the mitotic spindle through their kinetochores. The checkpoint signal originates from unattached kinetochores, where there is an enrichment of SAC proteins. Whether the enrichment of all SAC proteins is crucial for SAC signaling is unclear. Here, we provide evidence that, in fission yeast, recruitment of the kinase Mph1 is of vital importance for a stable SAC arrest. An Mph1 mutant that eliminates kinetochore enrichment abolishes SAC signaling, whereas forced recruitment of this mutant to kinetochores restores SAC signaling. In bub3Δ cells, the SAC is functional when only Mph1 and the Aurora kinase Ark1, but no other SAC proteins, are enriched at kinetochores. We analyzed the network of dependencies for SAC protein localization to kinetochores and identify a three-layered hierarchy with Ark1 and Mph1 on top, Bub1 and Bub3 in the middle, and Mad3 as well as the Mad1-Mad2 complex at the lower end of the hierarchy. If Mph1 is artificially recruited to kinetochores, Ark1 becomes dispensable for SAC activity. Our results highlight the crucial role of Mph1 at kinetochores and suggest that the Mad1-Mad2 complex does not necessarily need to be enriched at kinetochores for functional SAC signaling.

German adaptation of the Resources for Enhancing Alzheimer's Caregiver Health II: study protocol of a single-centred, randomised controlled trial.

BACKGROUND: Caring for a family member with dementia is extremely stressful, and contributes to psychiatric and physical illness among caregivers. Therefore, a comprehensive programme called Resources for Enhancing Alzheimer's Caregiver Health II (REACH II) was developed in the United States to enhance the health of Alzheimer's caregivers. REACH II causes a clear reduction of the stress and burdens faced by informal caregivers at home. The aim of this protocol is to adapt, apply, and evaluate this proven intervention programme in a German-speaking area for the first time. This newly adapted intervention is called Deutsche Adaption der Resources for Enhancing Alzheimer's Caregiver Health (DeREACH). METHODS: A total of 138 informal caregivers at home are recruited in a single-centred, randomised controlled trial. The intervention (DeREACH) consists of nine home visits and three telephone contacts over six months, all of which focus on safety, psychological well-being and self-care, social support, problem behaviour and preventive health-related behaviours. A complex intervention assessment on effectiveness will be adopted when the primary outcome - namely, the reduction of caregiver burden - and other secondary outcomes, including changes with regard to anxiety and depression, somatisation, health-related quality of life, and perceived social support, are measured at baseline, as well as immediately and three months after the intervention. The change from baseline to post-intervention assessment with regard to the primary outcome will be compared between treatment and control group using t-tests for independent samples. DISCUSSION: It is anticipated that this study will show that DeREACH effectively reduces caregiver burden and therefore works under the conditions of a local German health-care system. If successful, this programme will provide an effective intervention programme in the German-speaking area to identify and develop the personal capabilities of informal caregivers to cope with the burdens of caring for people with dementia.

Glucose stress causes mRNA retention in nuclear Nab2 condensates.

Nuclear mRNA export via nuclear pore complexes is an essential step in eukaryotic gene expression. Although factors involved in mRNA transport have been characterized, a comprehensive mechanistic understanding of this process and its regulation is lacking. Here, we use single-RNA imaging in yeast to show that cells use mRNA retention to control mRNA export during stress. We demonstrate that, upon glucose withdrawal, the essential RNA-binding factor Nab2 forms RNA-dependent condensate-like structures in the nucleus. This coincides with a reduced abundance of the DEAD-box ATPase Dbp5 at the nuclear pore. Depleting Dbp5, and consequently blocking mRNA export, is necessary and sufficient to trigger Nab2 condensation. The state of Nab2 condensation influences the extent of nuclear mRNA accumulation and can be recapitulated in vitro, where Nab2 forms RNA-dependent liquid droplets. We hypothesize that cells use condensation to regulate mRNA export and control gene expression during stress.

Probing Liquid-Liquid Phase Separation of RNA-Binding Proteins In Vitro and In Vivo.

Biomolecular condensates and the concept of liquid-liquid phase separation (LLPS) have transformed cell biology in recent years. Condensates organize cellular content and compartmentalize biochemical reactions, in particular many processes involving RNA. This protocol is aimed at readers new to the LLPS field who want to test their protein or cellular structure of interest. We describe the basic principles of liquid-liquid phase separation, and outline initial approaches-both in vitro and in yeast cells-for the characterization of a candidate cellular condensate. First, we focus on strategies to purify phase-separating proteins and to reconstitute condensates from recombinant proteins in vitro for observation by light microscopy. Second, we describe in vivo experiments (including fluorescence recovery after photobleaching (FRAP) microscopy and 1,6-Hexanediol treatment) to test whether a subcellular structure displays liquid-like behavior in cells.

Case management for people with dementia living at home and their informal caregivers: A scoping review.

Case management is a complex intervention aimed at addressing a variety of health needs of people in their social environment. Case management for people with dementia is often poorly defined and insufficiently described. The crucial process steps are often not well understood. We aim to map and compare the key components, processes and contextual factors of case management programmes for dementia and to explore aspects of the interventions' generalisability. Our search covered the databases PubMed, CINAHL, Cochrane and GeroLit, as well as policy papers from international organisations. We included qualitative, quantitative and mixed-methods studies in the English or German language that was published between 1999 and 2020. The programmes were analysed according to programme characteristics, case management intervention and the structural and processing conditions. We identified 67 studies dealing with 25 programmes. Approximately half of the programmes were investigated in randomised controlled trials, two programmes used a mixed-methods design and the remaining were the subject of pre-post cohort studies. Participants in the studies were predominantly dyads of people with dementia and their informal caregivers. About half of the programmes reported a theoretical framework. All the programmes were derived from case management approaches or referred to such approaches. Despite huge differences in implementation, all the programmes covered the case management steps. In 14 out of 25 programmes, case management was carried out without additional intervention, the other programmes provided mainly education and training for informal caregivers. Costs of the case management interventions were stated in more than half of the programmes.The effectiveness and generalisability of dementia-specific case management interventions could be enhanced if the framework introduced in the review was used in the future by policy, practice and research.